Selective targeting of matrix metalloproteinases for developing preterm labor therapeutics

项目摘要/摘要 我们已经证明基质金属蛋白酶家族的特定成员(基质金属蛋白酶-2和基质金属蛋白酶-9)是 在早产子宫肌层中过表达，这些酶加剧了子宫收缩反应 人类子宫平滑肌。因此，基质金属蛋白酶-2/9有可能作为阻断早产的治疗靶点。 金属蛋白酶组织抑制因子(TIMPs)是一类具有亚纳摩尔亲和力的内源性基质金属蛋白酶抑制剂。 考虑到MMPs在细胞功能中的多方面作用，我们的目标是针对特定的MMPs(MMP2/9)， 负责早产患者宫缩，具有高选择性，同时避免 与其他有益的MMP的相互作用。我们的中心假设是选择性TIMP蛋白 可以开发治疗方法来促进子宫静息。我们将使用蛋白质工程技术 例如定向进化以产生基于TIMP蛋白支架治疗早产的宫缩候选药物 劳力。在目标1中，我们将使用定向进化和酵母表面展示技术来设计基于TIMP的蛋白质 以提高对基质金属蛋白酶-2/9的结合选择性。目标2，我们将评估野生型和 经改造的TIMP可减少人类子宫组织的收缩。这些数据预计将是重要的 因为它们将为在临床前的早产儿体内模型中进行候选药物测试提供基础 劳力。

PROJECT SUMMARY/ABSTRACT We have shown that specific members of the matrix metalloproteinase family (MMP-2 and MMP-9) are overexpressed in preterm laboring myometrium and that these enzymes exacerbate contractile responses in human uterine smooth muscle. Therefore, MMP-2/9 may serve as therapeutic targets to block preterm labor. Tissue inhibitors of metalloproteinases (TIMPs) are endogenous MMP inhibitors with subnanomolar affinity. Considering multifaceted role of MMPs in cellular function, we aim to target specific MMPs (MMP-2/9), responsible for uterine contraction in patients with preterm labor, with high selectivity while avoiding interactions with other beneficial MMPs. Our central hypothesis is that selective TIMP protein-based therapeutics can be developed to promote uterine quiescence. We will use protein engineering techniques such as directed evolution to produce tocolytic drug candidates based on TIMP protein scaffold to treat preterm labor. In Aim 1, we will use directed evolution and yeast surface display to engineer TIMP-based protein scaffolds to improve binding selectivity toward MMP-2/9. In Aim 2, we will evaluate the ability of wild-type and engineered TIMPs to reduce contractions in human uterine tissue. These data are expected to be significant because they will provide the foundation for candidate drug testing in preclinical in vivo models of preterm labor.